酪蛋白糖巨肽对小鼠肠道双歧杆菌增殖水平的影响

发布时间：2018-11-28 10:48

【摘要】：采用实时荧光定量聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)分析不同剂量酪蛋白糖巨肽(casein glycomacropeptide,CGMP)对小鼠肠道双歧杆菌增殖水平的影响,揭示乳源CGMP是否有增殖肠道关键益生菌的功能。选用50只健康BALB/c小鼠,随机分为对照组,安慰组(灌胃生理盐水0.2 m L),乳源CGMP低、中、高剂量组(灌胃等体积不同质量浓度的乳源CGMP),灌胃周期为2周。于灌胃后第0、3、5、7、9、11、15天及停止灌胃后1周(第21天)采集的小鼠新鲜粪便,并进行粪便菌体基因组DNA抽提。依据双歧杆菌的16S rRNA基因序列设计5对种属特异性引物,以两歧双歧杆菌(Bifidobacterium bifidum)CICC6071的基因组DNA作为标准品,进行梯度稀释制作标准曲线;分析样品PCR扩增产物的熔解曲线,评价反应的特异性。结果表明:Real-time PCR可准确定量小鼠肠道内双歧杆菌数量;且灌胃中剂量乳源CGMP可以促进小鼠肠道内双歧杆菌的增殖,在灌胃第11天时小鼠肠道内双歧杆菌数量达到最高值,即乳源CGMP对小鼠肠道双歧杆菌的增殖作用存在剂量选择性。
[Abstract]:Real-time fluorescent quantitative polymerase chain reaction (real-time polymerase chain reaction,real-time PCR) was used to analyze the effects of different doses of casein glycosaminopeptide (casein glycomacropeptide,CGMP) on the proliferation of bifidobacterium in mice. To reveal whether milk source CGMP has the function of proliferating key probiotics in intestinal tract. Fifty healthy BALB/c mice were randomly divided into two groups: the placebo group (0.2 m L), by gastric administration of normal saline) had low milk-source CGMP, and the high dose group (intragastric instillation of milk source CGMP), of equal volume and different concentration) had a period of 2 weeks. Fresh feces were collected from mice on the 15th day after oral administration and 1 week (21 days) after gastric administration, and genomic DNA was extracted from the feces. According to the 16s rRNA gene sequence of Bifidobacterium, 5 pairs of species-specific primers were designed. The genomic DNA of Bifidobacterium bifidobacterium (Bifidobacterium bifidum) CICC6071 was used as standard material, and the standard curve was made by gradient dilution. The melting curve of PCR amplification products was analyzed and the specificity of the reaction was evaluated. The results showed that Real-time PCR could accurately quantify the number of bifidobacterium in the intestinal tract of mice. The medium dose CGMP could promote the proliferation of bifidobacterium in the intestinal tract of mice, and the number of bifidobacterium in the intestine of mice reached the highest value on the 11th day of gastric administration, that is, the dose selective effect of milk source CGMP on the proliferation of bifidobacterium in the intestinal tract of mice.
【作者单位】：天津市食品生物技术重点实验室天津商业大学生物技术与食品科学学院;
【基金】：国家自然科学基金面上项目(31071522)
【分类号】：R151

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